This chapter makes a comparative study between two kinds of biosurfactants (BSs) consisting of lipopeptides and glycoproteins. Although both BSs are produced by microorganisms, they have different properties and characteristics. Usually, glycoproteins are cell bound to the plasmatic membrane of bacteria, whereas lipopeptides are produced extracellularly presenting different schemes of separation and purification. The current chapter includes reviews of various methodologies proposed by different authors for the extraction and purification of glycoproteins and lipopeptides, ionic charge or the hydrophilic-lipophilic balance (HLB), which are of special criteria for cosmetic and pharmaceutical applications of BSs. Finally, a review on number of publications related to the production of lipopeptides and glycoproteins is included in this chapter.